LOCUS       sequence-411018-        5607 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISCysp13 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5607)
  AUTHORS   Xiang G, Li Y, Sun J, Huo Y, Cao S, Cao Y, Guo Y, Yang L, Cai Y, 
            Zhang YE, Wang H
  TITLE     Evolutionary mining and functional characterization of TnpB 
            nucleases identify efficient miniature genome editors.
  JOURNAL   Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x.
  PUBMED    37386294
REFERENCE   2  (bases 1 to 5607)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x."
COMMENT     Sequence Label: pBad33_ISCysp13_reRNA
FEATURES             Location/Qualifiers
     source          1..5607
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        19..53
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             69..80
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1653..1670)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1823..1909
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2001..2028
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2047..2138
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2596..3051
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2683..2702)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2893..2914
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3607..4266)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4181..4200
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4267..4369)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4761..4778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4895..5440)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(4975..4994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5516..5535
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgacagctta tcatcgatct gacagctagc tcagtcctag gtataatgct agccccggga
       61 aaagatctaa agaggagaaa gaattcatgg ctactaaaag aatcacgttt agactttacc
      121 caaataaaga acaaaatgag aaacttcatt attggagacg tttacacaaa gatttataca
      181 atgcttgtgt agttaatcga aaaactcaat acaaaaagtt tggtaaatcc attaattact
      241 ttgaccaaca aaactcatta cctgagttta aaaaagattg gccagaatac aaagaccttg
      301 gtagccatgc ccttcaagca actgtaaaac gggttgattt tgctttccaa cgttttttca
      361 agattaaatc aggttatccg aaactaaagt cttcaagaag atataaaggg tggacttatc
      421 ctgataaagc tggttggaaa atagacagta atggacatca tggaaagttg acactttcta
      481 attttggaac aataaagatg aggggtaaag caagagattg gggaacacca aatacttgca
      541 ctatcatctt taaacaagga aaatggtacg cttctgttac tgttagttgt gatccaaaac
      601 gagaaacagg taaagaggct tgtggtatcg attttgggac tcatcatgca cttgcttttg
      661 acgatggcac tgttattgat aatcctagat ttctcaaaca atcacaggat aaaatcaaca
      721 aaacagcaaa gaaatctaga aagaaacgtg caggaaaaaa gggagtaaaa gggtctagaa
      781 gatggaaaaa agctgttaga aaaatcggta agattcaatc tttagttggt cgtcaaagac
      841 aaaactggca tcatcaagtt tccacacaaa tagttagctg taatagcttg gtggcaactg
      901 agaaattaaa cctaaaagga atgaccaaaa aagctaaaag aggtagcaaa agaaaacgtc
      961 aaaagactgg attaaatcgg tcaattcttg acgttggaat tggtaactta aaatcattaa
     1021 taaaaagcaa agttaccgaa gcaggtgggt tttacattga ggttcctact cgaaaagtta
     1081 agccatctca gacttgtcct aattgtggtc atcaaaagaa aaagacatta gcagaacgcc
     1141 agcacaactg cgaaaagtgc ctttatagtt gcgacagaga tgttgcatca gcccaggtgt
     1201 gtttaaactg ggcaaggggg caggaactgt cctctttaga cgtagagtcg cctagctcta
     1261 ctttatgcgg aagcatgagg caactagggg cgaaaaagca caacaatgca aagaagcgtc
     1321 agaaacatct agctcaacaa agtggctgag agatctttga cagctagctc agtcctaggt
     1381 ataatagcct ttatagttgc gacagagatg ttgcatcagc ccaggtgtgt ttaaactggg
     1441 caagggggca ggaactgtcc tctttagacg tagagtcgcc tagctctact ttatgcggaa
     1501 gcatgaggca actaggggcg aaaaagcaca acaatgcaaa gaagcgtcag aaacatctag
     1561 ctcaacaaag tggctagatg tagttcatgc tcggagatca tcattgcgtt ttttaagctt
     1621 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     1681 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     1741 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     1801 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1861 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     1921 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     1981 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     2041 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2101 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2161 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2221 tggtgaaagt aaaagatgct gaagatcagt tgggtgcagc aaactattaa ctggcgaact
     2281 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2341 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2401 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2461 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2521 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2581 actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2641 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2701 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2761 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg
     2821 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2881 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcgggct
     2941 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     3001 tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt taaaaggatc
     3061 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3121 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3181 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3241 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     3301 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3361 cctacatacc tcgctctgct aatcctgtta ccagtcaggc atttgagaag cacacggtca
     3421 cactgcttcc ggtagtcaat aaaccggtaa accagcaata gacataagcg gctatttaac
     3481 gaccctgccc tgaaccgacg accgggtcga atttgctttc gaatttctgc cattcatccg
     3541 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     3601 aaaaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct
     3661 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     3721 cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat
     3781 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa
     3841 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     3901 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga
     3961 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     4021 cagctcaccg tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag
     4081 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     4141 cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     4201 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     4261 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     4321 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattt
     4381 tcgccaaaag ttggcccagg gcttcccggt atcaacaggg acaccaggat ttatttattc
     4441 tgcgaagtga tcttccgtca caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc
     4501 caacttactg atttagtgta tgatggtgtt tttgaggtgc tccagtggct tctgtttcta
     4561 tcagctgtcc ctcctgttca gctactgacg gggtggtgcg taacggcaaa agcaccgccg
     4621 gacatcagcg ctagcggagt gtatactggc ttactatgtt ggcactgatg agggtgtcag
     4681 tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga
     4741 tacaggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg
     4801 cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt
     4861 aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca
     4921 agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat
     4981 accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta
     5041 ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc
     5101 gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc
     5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca
     5221 ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta
     5281 aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt
     5341 caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt
     5401 ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca
     5461 gataaaatat ttctaggctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5521 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     5581 gtccggcgta gaggatctgc tcatgtt
//
